急性淋巴细胞白血病患儿骨髓间充质干细胞对K562/A02细胞株耐药的影响  

Effects of Acute Lymphoblastic Leukemia Children Bone Marrow Mesenchymal Stem Cells on Drug Resistance of K562/A02 Cell Line

作　　者：王昭霞[1] 杨志敏[1] 邹亚伟[2] 李敏敏[2] 陈福雄[2] 钟帼钰[2] 关镜明[2] 卫凤桂[2] 吴上志[2] 何振涛[2] 吴梓梁[2] 

机构地区：[1]威海市文登中心医院儿科,山东威海264400 [2]广州医学院第一附属医院儿科,广东广州510120

出　　处：《中国实验血液学杂志》2011年第1期19-23,共5页Journal of Experimental Hematology

基　　金：广东省医学科学技术研究基金(2005288);广州医学院博士启动基金(0706067)

摘　　要：本研究探讨急性淋巴细胞白血病患儿骨髓间充质干细胞(mesenchymal stem cells,MSC)对K562/A02细胞耐药性的影响及其机制。从白血病患儿骨髓中分离、培养并鉴定MSC;建立K562/A02细胞株与MSC共培养的体系,应用AnnexinV-FITC检测一定浓度的阿霉素(ADM)对不同培养条件下K562/A02细胞凋亡的影响;RT-PCR检测K562/A02细胞中的凋亡基因家族中bcl-2和bax基因;荧光定量PCR检测多药耐药基因mdr1浓度。结果表明,单独培养组白血病细胞早期凋亡率为(8.38±0.29)%,而黏附培养组为(1.97±0.11)%,差异具有统计学意义(p<0.05)。相对于单独悬浮培养的K562/A02细胞,黏附共培养的K562/A02细胞bcl-2基因表达明显增强,bcl-2/bax比值显著增高;K562/A02单独悬浮培养组、黏附共培养组细胞中mdr1的水平比较无统计学差异(p>0.05)。结论:白血病患儿MSC能够使白血病细胞逃避药物的促凋亡作用,K562/A02对阿霉素产生耐药性可能与黏附共培养后bcl-2基因表达增强有关,而与其本身含有的mdr1基因无关。The aim of study was to investigate the effect of acute lymphoblastic leukemia（ALL） children bone marrow mesenchymal stem cells（MSC） on resistance of K562/A02 cells and its mechanism.MSC obtained from bone marrow of AL children were cultured and identified.The co-culture of MSC and K562/A02 and the culture of K562/A02 cell suspention alone was performed,of which 2 kinds of cells were treated with same concentration of adriamycin（ADM）,and the rate of apoptosis was detected by flow cytometry,bcl-2 and bax of K562/A02 were detected by RT-PCR,while mdr1 gene level was detected by FQ-PCR.The results indicated that the MSC separation and proliferation were viable and steady.The apoptosis rate of the K562/A02 cells co-clutured with MSC was 1.97±0.11%,while apoptosis rate of the K562/A02 cells cultured alone was 8.38±0.29%,there was significant difference（p0.05）.As compared with the K562/A02 cells cultured alone,the bcl-2 gene expression in K562/A02 cells co-cultured with MSC obviously increased;ratio of bcl-2/bax was obviously enhanced.The mdr1 gene level in K562/A02 co-cultured with MSC was no statistical different from K562/A02 cultured alone（p0.05）,which suggested that adhesion co-cultured with MSC did not induce mdr1 expression higher than the culture of suspension.It is concluded that the MSC of ALL children can escape the leukemia cells from proapoptotic effect of drugs,the resistance of K562/A02 to ADM may be involved in enhancement of bcl-2 gene expression of K562/A02 cells co-cultured with MSC,but not in relation to mdr1 gene in K562/A02 cells themselves.

关 键 词：急性淋巴细胞白血病 间充质干细胞 K562/A02细胞 耐药 

分 类 号：R733.71[医药卫生—肿瘤]